CN105466870A - Cresol red colorimetric method for soil respiration measurement and measurement device - Google Patents
Cresol red colorimetric method for soil respiration measurement and measurement device Download PDFInfo
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- CN105466870A CN105466870A CN201510936974.5A CN201510936974A CN105466870A CN 105466870 A CN105466870 A CN 105466870A CN 201510936974 A CN201510936974 A CN 201510936974A CN 105466870 A CN105466870 A CN 105466870A
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- G—PHYSICS
- G01—MEASURING; TESTING
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- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
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Abstract
The invention discloses a cresol red colorimetric method for soil respiration measurement and a measurement device. The measurement device comprises a plastic test tube containing a cresol red reagent and a sealing cover. The plastic test tube and soil to be measured are sealed in the sealing cover, the absorbancy of the cresol red reagent is measured with the colorimetric method under the wavelength of 560 nm after respiration, and soil respiration intensity is measured through a standard curve. By preparing the cresol red reagent from the acid-base indicator cresol red, neutral salt and bicarbonate to absorb CO2 in soil, cultured soil respiration intensity is measured, and field soil respiration intensity can also be measured; strong base such as NaOH with high corrosivity is not needed, the reagent can be used repeatedly, sensitivity and safety performance are high, special or expensive instruments are not needed, the operation method is convenient and quick, and test data are reliable and accurate. The measurement device also has the advantages of being simple in structure, low in price, and convenient and rapid to use and operate.
Description
Technical field
The invention belongs to soil respiration determination techniques field, the cresol red colourimetry that specifically a kind of soil respiration measures and determinator.
Background technology
Soil respiration refers to that soil produces and to the process of air release of carbon dioxide.It comprises microbial respiratory, root system respiration and soil animal and breathes three bioprocess and an abiologic process: i.e. the chemical oxidation of gold of the soil organism under the high temperature conditions.Soil respiration is the important biomolecule index characterizing soil quality and fertility, and it reflects the intensity of biological activity of soil and soil material metabolism, can indicate process and the direction of ecosystem succession, is also directly affect Atmospheric CO
2concentration and change thereof, therefore measure soil respiration all significant in agricultural sciences, ecology and environmental science.Existing mode adopts highly basic that the corrosivity such as NaOH are strong to carry out the mensuration of soil respiration, experimental safe is poor, and reagent can only once use, substantially increase experimentation cost, and alkali liquor absorption titrimetry only amount of carbon dioxide be measure between 0.001g-0.002g comparatively accurate, measurement range is limited, and limitation is that the precision measured is undesirable.
Summary of the invention
The technical problem to be solved in the present invention is: the cresol red colourimetry providing a kind of soil respiration to measure and determinator, realize measuring culture soil to breathe, also can measure field soil to breathe, do not need the highly basic that the corrosivity such as NaOH are strong, and reagent is reusable, highly sensitive, without the need to special and expensive instrument and equipment, to solve down Problems existing in technology.
The technical scheme that the present invention takes is: the cresol red colourimetry that a kind of soil respiration measures, and its method comprises the following steps:
(1) solvent preparation: use acid base indicator cresol red and neutral salt and hydrocarbonate to be mixed with mix reagent, i.e. cresol red reagent;
(2) quantitative in aspiration step (1) cresol red reagent, in plastic test tube, builds lid;
(3) put into soil respiration determinator by after the plastic test tube uncap filling cresol red reagent in step (2), after being placed in soil surface to be measured sealing, starting timing, take out plastic test tube after 4-6h and cover lid;
(4) by the absorbance of the cresol red reagent colorimetric estimation cresol red reagent under 560nm wavelength after soil respiration in step (3), establishing criteria curve and blank calculate intensity of soil respiration.
Preferably, cresol red reagent comprises in every 1000mL water and puts into 0.0188g cresol red, 13.4160gKCl and 0.2520gNaHCO in above-mentioned steps (1)
3.
Preferably, above-mentioned steps (4) standard curve is: the reagent of use comprises the NaHCO of 0.1mol/L
3, the H of 0.2mol/L
2sO
4with the cresol red reagent in claim 2, concrete grammar step is: small beaker a) choosing 27 50mL, draws the NaHCO of the 0.1mol/L of 0mL, 1mL, 2mL, 3mL, 4mL, 5mL, 7.5mL, 10mL, 20mL respectively
3put into 9 small beakers wherein, and add distilled water or deionized water to 20mL; B) H of the 0.2mol/L of 20mL is added in another preparation 9 small beakers respectively
2sO
4; C) finally prepare all to put into the centrifuge tube that 1 fills 5mL cresol red reagent respectively in 9 small beakers, and uncap; D) NaHCO will be filled respectively
3, H
2sO
4put into packoff with each composition one group of the small beaker of the centrifuge tube of cresol red reagent to seal, total nine groups; E) then H will be filled across sealing bag
2sO
4small beaker in H
2sO
4pour into lentamente and fill NaHCO
3small beaker in, and start timing, after 4-6h, take out the centrifuge tube filling cresol red, and cover lid rapidly, colorimetric estimation absorbance under 560nm wavelength, the CO discharged with each small beaker
2amount be ordinate, with the absorbance of cresol red in centrifuge tube for horizontal ordinate, drawing standard curve, repeats 3 times.
Preferably, in above-mentioned steps (4), intensity of soil respiration comprises field soil respiratory intensity and cultivates respiratory intensity, and its computing formula is as follows:
In above formula: m
2-be field soil CO
2quality (
);
Respiratory area (the m that S-packoff contacts with field soil to be measured
2);
T
1-field soil breathing time (h);
M
2-be culture soil CO
2quality (
);
The quality of M-culture soil (
);
T
2-culture soil breathing time (h).
A kind of soil respiration determinator, comprise plastic test tube and seal closure one, described plastic test tube is fixedly connected on test tube rack, described test tube rack is plugged in soil to be measured, described seal closure one covers at soil surface to be measured, and placed wherein by plastic test tube, lower end arranges soil sealant in one week.
Preferably, above-mentioned test tube rack comprises employing steel wire and is arranged to spiral helicine socket part and steel wire spiral is dwindled into the bottom support portion of socket part, steel wire is extended into shank downwards around center by described bottom support portion, steel wire winding test tube is adopted to realize the support of test tube, structure is simple, equipment cost is low, convenient to operation, greatly improves testing equipment and prepares efficiency.
A kind of soil respiration determinator, comprise plastic test tube, containers of soil to be measured and seal closure two, described plastic test tube is placed in beaker, and described containers of soil to be measured and plastic test tube are placed in seal closure two and keep hermetically sealed.
Beneficial effect of the present invention: compared with prior art, the present invention is by being mixed with CO in cresol red agent absorbent soil by acid base indicator cresol red and neutral salt and hydrocarbonate
2realize measuring culture soil respiratory intensity, also field soil respiratory intensity can be measured, do not need the highly basic that the corrosivity such as NaOH are strong, and reagent is reusable, sensitivity and security high, without the need to special and expensive instrument and equipment, method of operating is convenient and swift, and test figure is reliably accurate, the feature that the equipment that the present invention adopts also has that structure is simple, low price, use are convenient to operation.
Accompanying drawing explanation
Fig. 1 is the field soil respiration measuring device structural representation that the present invention adopts;
Fig. 2 is the culture soil respiration measuring device structural representation that the present invention adopts;
Fig. 3 is canonical plotting of the present invention.
In figure: 1-plastic test tube, 2-seal closure one, 3-test tube rack, 4-soil horizon to be measured, 5-soil sealant, 6-socket part, 7-bottom support portion, 8-shank, 9-seal closure two, 10-culture soil container to be measured, 11-beaker.
Embodiment
Below in conjunction with accompanying drawing and specific embodiment, invention is described further.
Embodiment 1: plant the cresol red colourimetry that soil respiration measures, its method comprises the following steps:
(1) solvent preparation: use acid base indicator cresol red and neutral salt and hydrocarbonate to be mixed with mix reagent, i.e. cresol red reagent;
(2) quantitative in aspiration step (1) cresol red reagent, in plastic test tube, builds lid;
(3) put into soil respiration determinator by after the plastic test tube uncap filling cresol red reagent in step (2), after being placed in soil surface to be measured sealing, starting timing, take out plastic test tube after 4-6h and cover lid;
(4) by the absorbance of the cresol red reagent colorimetric estimation cresol red reagent under ultraviolet-visible spectrophotometer 560nm wavelength after soil respiration in step (3), establishing criteria curve and blank calculate intensity of soil respiration, intensity of soil respiration comprises field soil respiratory intensity and cultivates respiratory intensity, and its computing formula is as follows:
In above formula: m
2-be field soil CO
2quality (
);
Respiratory area (the m that S-packoff contacts with field soil to be measured
2);
T
1-field soil breathing time (h);
M
2-be culture soil CO
2quality (
);
The quality of M-culture soil (
);
T
2-culture soil breathing time (h).
Preferably, cresol red reagent comprises in every 1000mL water and puts into 0.0188g cresol red, 13.4160gKCl and 0.2520gNaHCO3 in above-mentioned steps (1).
Preferably, above-mentioned culture soil is: the pedotheque fetched from field is placed in spacious calm free of contamination laboratory, be placed on plastic paper by as far as possible uncovered for soil tiling, stir every day once, air-dry 7 days, the moisture in soil is evaporated completely, fetch after 7 days and grind in porcelain mill alms bowl, foreign material in soil are taken out, as dry branches and fallen leaves, rubbish incidental handstone etc.Then slowly grind, make the whole sieve crossing 2mm of soil sample fetched, ground later soil sample put into label in sealing bag and properly preserve, take 5g soil sample during test and put into 50ml small beaker, add 5ml distilled water, seal the opening of small beaker with preservative film and pinion guarantee sealing with bungee, the small beaker handled well being placed in the incubator of 25 DEG C and cultivating, taking out after 48h and be culture soil.
During test, the small beaker filling culture soil is put in large sealing bag and seal up opening and put dark local black out into and breathe, take out after 6h and cover rapidly centrifuge tube lid and cresol red reagent to be poured in cuvette and first measure under wavelength is 560nm with being placed on ultraviolet-visible spectrophotometer after reagent rinse one time again with distilled water washing.
Preferably, above-mentioned steps (4) standard curve is: the reagent of use comprises the NaHCO of 0.1mol/L
3, the H of 0.2mol/L
2sO
4with the cresol red reagent in claim 2, concrete grammar step is: small beaker a) choosing 27 50mL, draws the NaHCO of the 0.1mol/L of 0mL, 1mL, 2mL, 3mL, 4mL, 5mL, 7.5mL, 10mL, 20mL respectively
3put into 9 small beakers wherein, and add distilled water or deionized water to 20mL; B) H of the 0.2mol/L of 20mL is added in another preparation 9 small beakers respectively
2sO
4; C) finally prepare all to put into the centrifuge tube that 1 fills 5mL cresol red reagent respectively in 9 small beakers, and uncap; D) NaHCO will be filled respectively
3, H
2sO
4put into packoff with each composition one group of the small beaker of the centrifuge tube of cresol red reagent to seal, total nine groups; E) then H will be filled across sealing bag
2sO
4small beaker in H
2sO
4pour into lentamente and fill NaHCO
3small beaker in, and start timing, after 4-6h, take out the centrifuge tube filling cresol red, and cover lid rapidly, colorimetric estimation absorbance under 560nm wavelength, the CO discharged with each small beaker
2amount be ordinate, with the absorbance of cresol red in centrifuge tube for horizontal ordinate, drawing standard curve, repeat 3 times, gained typical curve is as shown in Figure 3.
Cresol red colourimetry only needs making typical curve to carry out reading, just eliminates the concentration determination of alkali lye, the concentration determination of acid, also has the calculating of complexity during titration.Be not only in measurement result, alkali liquor absorption titrimetry is more numerous and diverse than cresol red colourimetry in operability.The error using cresol red colourimetry also to eliminate NaOH solution and H2SO4 solution to produce in process for preparation, also eliminate in measurement and will use the loaded down with trivial details of titrimetry and the upper error produced of operation, cresol red colourimetry just directly can be carried out machine measurement after absorption carbon dioxide on ultraviolet-visible spectrophotometer, substantially increase work efficiency to it also avoid in measurement transfer and think the error produced, cresol red solution can recycling use, without waste, after measuring, unspent cresol red solution again can from new use after placing, because cresol red and carbon dioxide reaction just create physical change, and use alkali liquor absorption titrimetry to carry out mensuration to create chemical change and just cannot again utilize, the angle of environmental protection, cresol red colourimetry is also save, not extravagant and wasteful, meet the principle of modern environmental protection and saving experiment.
Embodiment 2: a kind of soil respiration determinator, described plastic test tube 1 and seal closure 1, described plastic test tube 1 is fixedly connected on test tube rack 3, described test tube rack 3 is plugged in soil 4 to be measured, described seal closure 1 covers on soil 4 surface to be measured, plastic test tube 1 is placed wherein, lower end arranges soil sealant 5 in one week, seal closure 1 adopts stainless steel cup, test tube rack 3 comprises employing steel wire and is arranged to spiral helicine socket part 6 and steel wire spiral is dwindled into the bottom support portion 7 of socket part, steel wire is extended into shank 8 downwards around center by described bottom support portion 7, steel wire winding test tube is adopted to realize the support of test tube, structure is simple, equipment cost is low, convenient to operation, greatly improve testing equipment and prepare efficiency.
Embodiment 3: a kind of soil respiration determinator, described plastic test tube 1, containers of soil to be measured 10 and seal closure 29, described plastic test tube 1 is placed in beaker 11, described containers of soil to be measured 10 and plastic test tube 1 are placed in seal closure 29 and keep hermetically sealed, and seal closure 29 adopts 14# valve bag.
The above; be only the specific embodiment of the present invention; but protection scope of the present invention is not limited thereto; anyly be familiar with those skilled in the art in the technical scope that the present invention discloses; change can be expected easily or replace; all should be encompassed within protection scope of the present invention, therefore, protection scope of the present invention should be as the criterion with the protection domain of described claim.
Claims (7)
1. a cresol red colourimetry for soil respiration mensuration, is characterized in that: the method comprises the following steps:
(1) solvent preparation: use acid base indicator cresol red and neutral salt and hydrocarbonate to be mixed with mix reagent, i.e. cresol red reagent;
(2) quantitative in aspiration step (1) cresol red reagent, in plastic test tube, builds lid;
(3) put into soil respiration determinator by after the plastic test tube uncap filling cresol red reagent in step (2), after being placed in soil surface to be measured sealing, starting timing, take out plastic test tube after 4-6h and cover lid;
(4) by the absorbance of the cresol red reagent colorimetric estimation cresol red reagent under 560nm wavelength after soil respiration in step (3), establishing criteria curve and blank calculate intensity of soil respiration.
2. the cresol red colourimetry of a kind of soil respiration mensuration according to claim 1, is characterized in that: the middle cresol red reagent of described step (1) comprises in every 1000mL water puts into 0.0188g cresol red, 13.4160gKCl and 0.2520gNaHCO
3.
3. the cresol red colourimetry of a kind of soil respiration mensuration according to claim 1, is characterized in that: described step (4) standard curve is: the reagent of use comprises the NaHCO of 0.1mol/L
3, the H of 0.2mol/L
2sO
4with the cresol red reagent in claim 2, concrete grammar step is: small beaker a) choosing 27 50mL, draws the NaHCO of the 0.1mol/L of 0mL, 1mL, 2mL, 3mL, 4mL, 5mL, 7.5mL, 10mL, 20mL respectively
3put into 9 small beakers wherein, and add distilled water or deionized water to 20mL; B) H of the 0.2mol/L of 20mL is added in another preparation 9 small beakers respectively
2sO
4; C) finally prepare all to put into the centrifuge tube that 1 fills 5mL cresol red reagent respectively in 9 small beakers, and uncap; D) NaHCO will be filled respectively
3, H
2sO
4put into packoff with each composition one group of the small beaker of the centrifuge tube of cresol red reagent to seal, total nine groups; E) then H will be filled across sealing bag
2sO
4small beaker in H
2sO
4pour into lentamente and fill NaHCO
3small beaker in, and start timing, after 4-6h, take out the centrifuge tube filling cresol red, and cover lid rapidly, colorimetric estimation absorbance under 560nm wavelength, the CO discharged with each small beaker
2amount be ordinate, with the absorbance of cresol red in centrifuge tube for horizontal ordinate, drawing standard curve, repeats 3 times.
4. the cresol red colourimetry of a kind of soil respiration mensuration according to claim 1, is characterized in that: in described step (4), intensity of soil respiration comprises field soil respiratory intensity and cultivates respiratory intensity, and its computing formula is as follows:
In above formula: m
2-be field soil CO
2quality (
);
Respiratory area (the m that S-packoff contacts with field soil to be measured
2);
T
1-field soil breathing time (h);
M
2-be culture soil CO
2quality (
);
The quality of M-culture soil (
);
T
2-culture soil breathing time (h).
5. a soil respiration determinator, it is characterized in that: described plastic test tube (1) and seal closure one (2), described plastic test tube (1) is fixedly connected on test tube rack (3), described test tube rack (3) is plugged in soil to be measured (4), described seal closure one (2) covers on soil to be measured (4) surface, placed wherein by plastic test tube (1), lower end arranges soil sealant (5) for one week.
6. a kind of soil respiration determinator according to claim 1, it is characterized in that: described test tube rack (3) comprises employing steel wire and is arranged to spiral helicine socket part (6) and steel wire spiral dwindled into the bottom support portion (7) of socket part, and steel wire is extended into shank (8) downwards by described bottom support portion (7) around center.
7. a soil respiration determinator, it is characterized in that: described plastic test tube (1), containers of soil to be measured (9) and seal closure two (10), described plastic test tube (1) is placed in beaker (11), and described containers of soil to be measured (9) and plastic test tube (1) are placed into and keep hermetically sealed in seal closure two (10).
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Cited By (2)
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CN106525695A (en) * | 2017-01-10 | 2017-03-22 | 贵州大学 | Soil capillary porosity tester and testing method thereof |
CN110412239A (en) * | 2018-04-27 | 2019-11-05 | 大连理工大学 | A kind of quickly detection Microbial biomass C O2The gel test paper and its application method of respiratory rate |
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Cited By (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN106525695A (en) * | 2017-01-10 | 2017-03-22 | 贵州大学 | Soil capillary porosity tester and testing method thereof |
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CN110412239A (en) * | 2018-04-27 | 2019-11-05 | 大连理工大学 | A kind of quickly detection Microbial biomass C O2The gel test paper and its application method of respiratory rate |
CN110412239B (en) * | 2018-04-27 | 2021-08-10 | 大连理工大学 | Rapid detection of microbial CO2Gel test paper for respiratory rate and use method thereof |
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Application publication date: 20160406 |